聚酰胺-胺型树状高聚物介导短发卡RNA转染小鼠腹膜组织的实验研究

Transfer of plasmid expressing short hair RNA into mouse peritoneum mediated by polyamidoamine dendrimers

目的研究聚酰胺-胺型树状高聚物(PAMAM)载体介导pHK-shRNA质粒在小鼠腹膜组织的表达情况。方法将昆明小鼠随机分为转染组和对照组,分别经腹腔注射PAMAM G9/pHK-shRNA复合物或生理盐水,于24h、48h、96h和168h留取壁层和脏层腹膜组织,在荧光显微镜下观察壁层腹膜绿色荧光蛋白(GFP)的表达,同时采用Western Blot检测脏层腹膜GFP的表达。结果正常对照组的小鼠腹膜组织未能检测到GFP表达,而PAMAM G9/pHK-shRNA转染组的各时间点均可检测到GFP表达,以48h表达最强,显著高于其他各时间点(P<0.01)。Western Blot结果与冰冻组织切片的结果一致。结论PAMAM载体可介导pHK-shRNA转染正常小鼠的腹膜组织,为进一步体内研究提供实验依据。

Objective To investigate the expression of pHK-shRNA plasmid mediated by polyamidoamine dendrimers（PAMAM）in mouse peritoneum. Methods Kunming mice were randomly divided into transfection group and control group, and were intraperitoneally injected with PAMAM G9/pHK-shRNA complex and saline, respectively. Parietal and visceral peritoneum were collected after the injection for 24h, 48h, 96h and 7d, and the expression of green fluorescent protein （GFP） in peritoneum was detected by fluorescence microscope and western blot. Results Expression of GFP was not detected in control group, and was detected in peritoneum of transfection group at all time points. Western blot and observation of tissue sections showed that the peak of the expression was at 48h after the transfection. Conclusion pHK-shRNA plasmid can be efficiently transferred into mouse peritoneal tissue by PAMAM, which provides an approach for in vivo experimental studies.